Methods for sequencing a biomolecule by detecting relative positions of hybridized probes
First Claim
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1. A method for determining a sequence of a biomolecule, the method comprising the steps of:
- (a) identifying a set of k−
1-length subsequences that represent a plurality of substrings of a sequence string s of the biomolecule;
(b) for each of the k−
1-length subsequences, forming a pool of probes comprising four different k-mer extensions of the k−
1-length subsequence;
(c) for each pool formed in step (b);
(i) hybridizing the biomolecule with the four k-mer probes making up the pool; and
(ii) detecting relative positions of the k-mer probes that have attached to the biomolecule; and
(d) ordering the subsequences corresponding to the detected attached probes to determine the sequence string s of the biomolecule.
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Abstract
A sequencing method is presented in which a biomolecule is hybridized with a specially chosen pool of different probes of known sequence which can be electrically distinguished. The different probe types are tagged such that they can be distinguished from each other in a Hybridization Assisted Nanopore Sequencing (HANS) detection system, and their relative positions on the biomolecule can be determined as the biomolecule passes through a pore or channel. The methods eliminate, resolve, or greatly reduce ambiguities encountered in previous sequencing methods.
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Citations
12 Claims
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1. A method for determining a sequence of a biomolecule, the method comprising the steps of:
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(a) identifying a set of k−
1-length subsequences that represent a plurality of substrings of a sequence string s of the biomolecule;(b) for each of the k−
1-length subsequences, forming a pool of probes comprising four different k-mer extensions of the k−
1-length subsequence;(c) for each pool formed in step (b); (i) hybridizing the biomolecule with the four k-mer probes making up the pool; and (ii) detecting relative positions of the k-mer probes that have attached to the biomolecule; and (d) ordering the subsequences corresponding to the detected attached probes to determine the sequence string s of the biomolecule. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification